Retinal Pigment Epithelium (RPE)

Retinal Pigment Epithelium Model Highlights

Accelerate your lead compound selection by understanding their mode of action in functional retinal tissue

Mature and functional RPE model

Well-characterised and reproducible

Rapid evaluation of gene therapy vectors

A functional monolayer in vitro model of retinal pigment epithelial cells generated from human iPSCs for accurate prediction of clinical outcomes

The retinal pigment epithelial (RPE) cell model, used for our services, is composed of a monolayer of RPE cells. Uniquely, our RPE cells are derived from healthy donor iPSCs from the same genetic background as retinal organoids, allowing parallel assessment of both RPE and neurosensory retina. We also have the capability to generate RPE cells from customer-supplied iPSCs. The 24-well Transwell® plates format allows for flexibility in dosing and analytical readouts, including functional assessment of the cells. The RPE characterisation is extensive including morphology assessment, pigmentation, RPE-specific expression at the protein level (BEST1, TYRP1), analysis of phagocytosis of photoreceptor outer segments, trans-epithelial resistance (TEER), polarity of apical Pigment Epithelium-Derived Factors (PEDF) and basal vascular endothelial growth factor (VEGF) secretion.

Applications

Gene therapy including in vitro viral vector assessment
Disease modelling
Investigational drug safety and efficacy
Effect of a compounds on RPE barrier integrity, phagocytic function, viability, gene and protein expression, growth factor secretion

Available analytical readouts for services provided with RPE

Imaging
mRNA quantification by RT-qPCR
Transcriptomics by single-cell RNA sequencing (RNAseq)
Growth factor (VEGF, PEDF) secretion
Flow cytometry
Gene Expression
Effect on ultrastructure (Electron microscopy assessment)

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A microscope image — RPE cells displaying cobblestone morphology. Cells were immunolabeled with tight-junction ZO-1 marker (shown in green) and co-stained with nuclei marker, Hoechst (shown in blue).

Origin

Healthy donor

Outsourced

We do the work

Characteristics

Functional RPE

“The extensive scientific knowledge and rigor of Newcells’ staff in the field of ophthalmology as well as their client goal-oriented thinking allowed us to advance quickly in our proof of concept.” Dr Elke Vermassen, Sr. Product Development Manager, KiOmed Pharma

Model formats

RPE cells cultured in 24-well Transwell® plates

Cell Types

Retinal pigment epithelial (RPE) cells

Origin

Human iPSCs (healthy donor)

How to use our RPE model?

Outsource your experiments to us

Take advantage of our expertise: with a fast and reliable service, Newcells’ team designs the experiments with you to answer your questions. For example, gene therapy vector screening and efficacy testing can give you results in as little as 2-3 months. All bespoke projects are carried out in our state-of-the-art UK facilities.

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Applications for retinal pigment epithelial cells Close Open

Gene therapy including in vitro viral vector evaluation
Disease modelling
Investigational drug safety and efficacy
Mechanism of action of compound on the barrier

Role of RPE in the retina Close Open

The retinal pigment epithelium (RPE) is a single layer of hexagonal-shaped cells that is located between the photoreceptors of the retina and the blood vessels of the choroid, at the back of the eye. The RPE cell layer is critical to vision. The RPE reduces light damage in the eye, regulates the movement of nutrients and waste products in and out of the retina and maintains blood vessels by secreting growth factors. RPE is also responsible for the turnover of photoreceptors.

The RPE is crucial to vision, thus it is important to assess how it is affected by new therapeutic drugs, a task that can be performed in vitro.

Retinal pigment epithelial cells characterisation Close Open

In vitro retinal pigment epithelial cells are generated from human iPSCs to form a 2D monolayer. These RPE cells display typical cobblestone morphology and are pigmented.

Hexagonal morphology and pigmentation typically observed in mature RPE cells

The in vitro model closely recapitulates key functions found in vivo, including formation of tight barriers and phagocytosis of photoreceptor outer segments.

TEER measurements were carried out starting 7 days after seeding on the inserts. RPE reach maturation 4-6 weeks post-isolation when TEER starts to plateau.

Flow cytometry data illustrates a highly pure, differentiated population as demonstrated by high levels of expression of RPE markers BEST1, TYRP1 and PMEL17

Expression of RPE markers, TYRP1, PMEL17 and BEST1

Protein expression of RPE cells using flow cytometry after TEER plateaued. PMEL17 – a protein which is expressed in melanosome precursors (>95% expression). TYRP1 – expressed in mature RPE and is located in melanosomes (>85% expression). BEST1 – is a Ca2+-regulated chloride channel and is a critical for normal phagocytotic function (>98% expression)

Retinal pigment epithelial cells description Close Open

Cell types: RPE cells
Format: Monolayer, cultured in 24-well Transwell® plates
Main characteristics: Cobblestone morphology, pigmentation, express RPE cells biomarkers e.g., BEST1, TYRP1 and RPE65
Other characteristics: Formation of tight barrier and ability to phagocytose photoreceptor outer segments

RPE cells origin: healthy, patient-derived or gene edited cells Close Open

Our RPE is differentiated from fully-characterised human iPSCs from a heathy donor.

RPE can be generated from our clients’ iPSCs ( patient derived or gene -edited). Newcells also reprograms PBMCs and fibroblasts prior to differentiation into RPE.

Mechanism of action of compound on the RPE cell layer Close Open

The RPE model is ideal to assess the effect of compounds on the epithelium cell layer. Following the addition of a compound, the following physiological functions can be assessed:

barrier integrity
phagocytic function
cell viability
protein expression
growth factor secretion

The RPE service includes a set of functional assays including gene expression, RNAseq and imaging to determine the effect of drugs on epithelium and cellular structures.

Catalogue reference Close Open

Model	Sku no.	Format	Species	Readouts
RPE disease modelling	RSD0000RPE	24-well Transwell	Human	Brightfield imaging, Cell viability (ATP/LDH), Quantitative IF, Gene expression, Transmission (TEM) and Scanning (SEM) electron microscopy (optional), Phagocytosis of photoreceptor outer segment (optional)
RPE gene therapy assessment	RSG0000RPE	24-well Transwell	Human	Transduction efficiency of viral vector, Cell viability assessment (ATP/LDH), Therapy efficacy using IF and Gene expression

Test your novel therapeutics on two retina models :
RPE and organoids

Outsource experiments to us

We can test your compounds on both retinal organoids and RPE cells. This is particularly relevant for viral vector evaluation and disease modelling.

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Gene Therapy

Rapid in vitro preclinical evaluation of viral vectors for retinal therapy development.

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Disease Modelling

We ethically source patient lines or use gene edited lines from our customers to produce models of retinopathies.

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RPE differentiation

iPSCs, PBMCs and fibroblasts (patient derived or gene edited) supplied by our clients can be used as starting material to differentiate into RPE.

Outsource your experiments to us

Interested in our RPE model? Speak to one of our experts about your requirements.

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Retinal Pigment Epithelium (RPE)